Increase In Metabolite Levels Research Articles

Biomonitoring of Hydroxylated Polycyclic Aromatic Hydrocarbon Metabolites in Workers at a Waste-to-Energy Incinerator, Turin, Italy

This paper presents the results of the human biomonitoring of ten urinary OH-PAHs (hydroxylated polycyclic aromatic hydrocarbon) in a cohort of workers at an incinerator in Turin, Italy. Long-term exposure was assessed through repeated measurements at three time points: before the startup (T0), after 1 year (T1), and after 3 years (T2). Paired data were available for 26 subjects, seven administrative workers (AWs) and 19 plant workers (PWs). Short-term exposure was assessed by comparing start-end shift measurements. Due to the non-normal distribution of the data, the nonparametric Cuzick’s test for trend and the Wilcoxon signed-rank test for paired samples were used. Neither the trend nor the T0-T2 comparison tests resulted in statistically significant outputs in the two groups (q-value > 0.05), even when controlling for smoking habits. In relation to PWs, some of the metabolites were higher at T2 with respect to T0, but no linear increase was found. Conversely, 1-OH-PYR (ng/g creatinine) showed lower median values at T1 (61.5) and T2 (67) compared to the baseline (151.3). Similarly, short-term comparisons yielded no significant results, with rather overlapping distributions of values. Overall, no significant increases in metabolite levels were detected as a result of occupational exposure in the incinerator workers considered. These findings align with previous results for metals and ambient air measurements.

Prickly Ash Seeds Improve the Ruminal Epithelial Development and Growth Performance of Hu Sheep by Modulating the Rumen Microbiota and Metabolome.

It is known that the addition of feed rich in bioactive components to animal diets will affect rumen fermentation parameters and flora structure. However, research on the regulatory effects of prickly ash seeds (PASs) during rumen development or on the rumen microbiome and its metabolites in sheep is limited. The current study was designed to explore the effects of PASs on sheep rumen development and growth performance using metagenomics and metabolomics. Eighteen 3-month-old Hu lambs were randomly allotted to three different dietary treatment groups: 0% (basal diet, CK), 3% (CK with 3% PAS, low-dose PAS, LPS), and 6% (CK with 6% PAS, high-dose PAS, HPS) PASs. The lambs were slaughtered to evaluate production performance. Our results showed that dietary PAS addition improved the average daily gain and reduced the F/G ratio of the experimental animals. Additionally, the height and width of the rumen papilla in the treatment groups were significantly higher than those in the CK group. The fermentation parameters showed that the levels of acetate and butyrate were significantly higher in the LPS group than in the CK and HPS groups. The propionate levels in the HPS group were significantly higher than those in the CK and LPS groups. Metagenomics analysis revealed that PAS dietary supplementation improved the abundance of Clostridiales and Bacteroidales and reduced the abundance of Prevotella, Butyrivibrio, and Methanococcus. Metabolomic analyses revealed that increased metabolite levels, such as those of serotonin, L-isoleucine, and L-valine, were closely related to growth-related metabolic pathways. The correlations analyzed showed that papilla height and muscular thickness were positively and negatively correlated with serotonin and L-valine, respectively. Average daily gain (ADG) was positively and negatively correlated with L-valine and several Prevotella, respectively. In addition, muscular thickness was positively correlated with Sodaliphilus pleomorphus, four Prevotella strains, Sarcina_sp_DSM_11001, and Methanobrevibacter_thaueri. Overall, PAS addition improved sheep growth performance by regulating beneficial microorganism and metabolite abundances, facilitating bacterial and viral invasion resistance.

Enhanced Production of Therapeutic Metabolites in Cork-Oak Somatic Embryos under Abiotic Stress Conditions

This study investigates the enhanced production of key therapeutic metabolites (ellagic acid, serotonin, and chlorogenic acid) in response to abiotic stress in in vitro cultures of Quercus suber somatic embryos. Findings indicate significant increases in metabolite levels under various stress conditions, highlighting the potential for commercial-scale production of these compounds, known for their antioxidant, anticancer, and anti-COVID-19 properties. Under osmotic/saline stress, ellagic acid production significantly increased, representing an 80% increase compared to control conditions. In embryos exposed to different stressors, serotonin accumulation showed a six-fold increase under osmotic/saline stress. Although the elicitors used did not increase chlorogenic acid levels, exploring alternative stress types may enhance its production. This research paves the way for sustainable, large-scale production of health-beneficial metabolites, addressing global health challenges and promoting resource sustainability.

SrHPO4-coated Mg alloy implant attenuates postoperative pain by suppressing osteoclast-induced sensory innervation in osteoporotic fractures

Osteoporotic fractures have become a common public health problem and are usually accompanied by chronic pain. Mg and Mg-based alloys are considered the next-generation orthopedic implants for their excellent osteogenic inductivity, biocompatibility, and biodegradability. However, Mg-based alloy can initiate aberrant activation of osteoclasts and modulate sensory innervation into bone callus resulting in postoperative pain at the sequential stage of osteoporotic fracture healing. Its mechanism is going to be investigated. Strontium hydrogen phosphate (SrHPO4) coating to delay the Mg-based alloy degradation, can reduce the osteoclast formation and inhibit the growth of sensory nerves into bone callus, dorsal root ganglion hyperexcitability, and pain hypersensitivity at the early stage. Liquid chromatography-mass spectrometry (LC-MS) metabolomics analysis of bone marrow-derived macrophages (BMMs) treated with SrHPO4-coated Mg alloy extracts shows the potential effect of increased metabolite levels of AICAR (an activator of the AMPK pathway). We demonstrate a possible modulated secretion of AICAR and osteoclast differentiation from BMMs, which inhibits sensory innervation and postoperative pain through the AMPK/mTORc1/S6K pathway. Importantly, supplementing with AICAR in Mg-activated osteoclasts attenuates postoperative pain. These results suggest that Mg-induced postoperative pain is related to the osteoclastogenesis and sensory innervation at the early stage in the osteoporotic fractures and the SrHPO4 coating on Mg-based alloys can reduce the pain by upregulating AICAR secretion from BMMs or preosteoclasts.

The immediate metabolomic effects of whole-genome duplication in the greater duckweed, Spirodela polyrhiza.

In plants, whole-genome duplication (WGD) is a common mutation with profound evolutionary potential. Given the costs associated with a superfluous genome copy, polyploid establishment is enigmatic. However, in the right environment, immediate phenotypic changes following WGD can facilitate establishment. Metabolite abundances are the direct output of the cell's regulatory network and determine much of the impact of environmental and genetic change on the phenotype. While it is well known that an increase in the bulk amount of genetic material can increase cell size, the impact of gene dosage multiplication on the metabolome remains largely unknown. We used untargeted metabolomics on four genetically distinct diploid-neoautotetraploid pairs of the greater duckweed, Spirodela polyrhiza, to investigate how WGD affects metabolite abundances per cell and per biomass. Autopolyploidy increased metabolite levels per cell, but the response of individual metabolites varied considerably. However, the impact on metabolite level per biomass was restricted because the increased cell size reduced the metabolite concentration per cell. Nevertheless, we detected both quantitative and qualitative effects of WGD on the metabolome. Many effects were strain-specific, but some were shared by all four strains. The nature and impact of metabolic changes after WGD depended strongly on the genotype. Dosage effects have the potential to alter the plant metabolome qualitatively and quantitatively, but were largely balanced out by the reduction in metabolite concentration due to an increase in cell size in this species.

Abstract TP236: Temporal Metabolomic Profiling Exposed Affiliated Metabolites Altered And Associations To Outcome After Different Acute Brain Injuries

Background: Neuronal recovering processes after acute brain injuries such as aneurysmal subarachnoid hemorrhage (aSAH), ischemic stroke (IS) and traumatic brain injury (TBI) are complex including brain plasticity and synaptic regeneration mechanisms. Hypotheses: Transient changes in metabolome after different acute brain injuries may reveal affiliated metabolites, metabolic pathways and associations that can also reflect important biological mechanisms. Measurable temporal changes in serum metabolomic profile may be similar irrespective of the type of the brain injury. These metabolomic commonalities may be associated to outcome. Methods: Prospective cohort (n=74) consisted of IS (n=30), aSAH (n=31) and TBI (n=13) patients. Serum samples were collected in two time points after the insult (early 24-48h and late 120-192h). Orbitrap Profiling of 462 metabolites was used to measure metabolites. Outcome was measured 90 days after injury (mRS favorable 0-3, unfavorable 4-6). The metabolomics data was imputed for missing value using KNN with 20% cut off for missing value, log-transformed and autoscaled. For statistics, t-test and ANOVA was performed and FDR-corrected p-value 0.05 was considered to be statistically significant. Results: We identified four similarly temporally increased metabolite levels across all acute brain injury types (p<0.05, FDR corrected). Twenty-eight metabolites were shown to have similar trend when only IS and TBI groups were compared. Whereas outcome analysis showed three metabolites associating both favorable and unfavorable outcome across all acute brain injury types (p<0,05, FDR corrected). IS and TBI groups combined twenty common metabolites were identified to associate favorable and twenty-three with unfavorable outcome (p<0,05, FDR corrected). Conclusion: Similar metabolic changes across time were identified with apparent associations to outcome. This suggests important common metabolic characteristics irrespective of the type of brain insult and encourage further studies and validation. Identified metabolites may be further developed as a prognostic biomarker and may indicate new metabolic pathways active irrespective the type of brain injury reflecting biological mechanisms.

Abstract P030: Prospective analysis of plasma metabolites for early detection of high-grade serous ovarian cancer

Abstract Background: Ovarian cancer has poor prognosis due to >60% of the patients being diagnosed at late-stage. However, there is currently no ovarian cancer biomarker appropriate for screening. This may be, in part, due to many prior studies using clinical samples obtained at the time of diagnosis for biomarker discovery. Here, we sought to discover novel plasma metabolomic biomarkers for ovarian cancer early detection using prospectively collected blood samples. Methods: We examined 250 plasma metabolites in 93 high-grade serous ovarian cancer (HGSOC) cases with blood drawn up to five years prior to diagnosis and 93 matched controls in the prospective Nurses’ Health Studies and Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial. We also measured metabolites in 40 early-stage HGSOC cases with blood collected at diagnosis and 40 matched controls in a hospital-based study. We used conditional logistic regression to calculate the odds ratios (ORs) and 95% confidence intervals (CIs) of the association between one standard deviation increase in metabolite levels and ovarian cancer risk, and used metabolite set enrichment analysis (MSEA) to identify metabolite group associations. False discovery rate (FDR) was used to correct for multiple testing. For the prospectively collected blood samples, we conducted analyses by time between blood draw and diagnosis. Results: Plasma metabolites associated with ovarian cancer in blood samples collected prospectively were different than in blood samples collected at diagnosis of early-stage disease compared to controls. There were 45 individual metabolites associated with early-stage disease (FDR<0.05), and none of these metabolites were associated with ovarian cancer in the prospectively collected blood samples (FDR<0.05). There were 10 metabolites associated with ovarian cancer diagnosed within 3 years of blood collection (p<0.05), including 7 lipid metabolites (2 lysophosphatidylcholines, 2 diglycerides, 1 phosphatidylcholine, 1 cholesteryl esters, 1 phosphatidylethanolamines) that were inversely associated with ORs ranging from 0.61-0.66. Interestingly, the direction of association remained similar when restricting to cases with blood collected up to 2 years and 1 year prior to diagnosis, and the magnitude of association generally became stronger for cases closer to diagnosis. Similarly, distinct profiles were observed in MSEA, with diglycerides being inversely associated with ovarian cancer diagnosis in prospectively collected blood samples (FDR ranging from <0.05 to <0.20) and positively associated in blood collected at early-stage disease diagnosis (FDR<0.05).Conclusion: Our results revealed plasma metabolomic profiles differ between prospectively collected blood samples up to 3 years prior to diagnosis and blood samples collected at time of diagnosis, even for early-stage disease, suggesting the use of prospectively collected blood samples may discover promising novel early detection biomarkers for ovarian cancer. Citation Format: Naoko Sasamoto, Oana A. Zeleznik, Allison F. Vitonis, Daniel W. Cramer, Julian Avila-Pacheco, Clary B. Clish, Britton Trabert, Shelley S. Tworoger, Kathryn L. Terry. Prospective analysis of plasma metabolites for early detection of high-grade serous ovarian cancer. [abstract]. In: Proceedings of the AACR Special Conference: Precision Prevention, Early Detection, and Interception of Cancer; 2022 Nov 17-19; Austin, TX. Philadelphia (PA): AACR; Can Prev Res 2023;16(1 Suppl): Abstract nr P030.

Assessment of susceptibility to phthalate and DINCH exposure through CYP and UGT single nucleotide polymorphisms

Single nucleotide polymorphisms (SNPs) of cytochrome P450 (CYPs) and UDP-glucuronosyltransferase (UGTs) genes have been proposed to influence phthalates and 1,2-cyclo-hexanedicarboxylic acid diisononyl ester (DINCH) biotransformation but have not been investigated on a populational level. We investigated the role of SNPs in CYP2C9, CYP2C19, CYP2D6, UGT2B15, and UGT1A7 genes in the biotransformation of phthalates (DEHP, DEP, DiBP, DnBP, BBzP, DiNP, DidP) and DINCH by determining their urine metabolites. From the Slovenian study population of 274 men and 289 lactating primiparous women we obtained data on phthalate and DINCH urine metabolite levels (MEHP, 5OH-MEHP, 5oxo-MEHP, 5cx-MEPP, MEP, MiBP, MnBP, MBzP, cx-MINP, OH-MiDP, MCHP, MnPeP, MnOP, 5OH-MINCH, 5oxo-MINCH), SNP genotypes (rs1057910=CYP2C9*3, rs1799853=CYP2C9*2, rs4244285=CYP2C19*2, rs12248560=CYP2C19*17, rs3892097=CYP2D6*4, rs1902023=UGT2B15*2, and rs11692021=UGT1A7*3) and questionnaires. Associations of SNPs with levels of metabolites and their ratios were assessed by multiple linear regression and ordinary logistic regression analyses. Significant associations were observed for CYP2C9*2, CYP2C9*3, CYP2C19*17, and UGT1A7*3 SNPs. The most pronounced was the influence of CYP2C9*2 and *3 on the reduced DEHP biotransformation, with lower levels of metabolites and their ratios in men and women. In contrast, carriers of CYP2C19*17 showed higher urine levels of DEHP metabolites in both genders, and in women also in higher DiNP, DiDP, and DINCH metabolite levels. The presence of UGT1A7*3 was associated with increased metabolite levels of DINCH in men and of DiBP and DBzP in women. Statistical models explained up to 27% of variability in metabolite levels or their ratios. Our observations confirm the effect of CYP2C9*2 and *3 SNPs towards reduced DEHP biotransformation. We show that CYP2C9*2, CYP2C9*3, CYP2C19*17, and UGT1A7*3 SNPs might represent biomarkers of susceptibility or resilience in phthalates and DINCH exposure that have been so far unrecognised.

Primary Metabolite Profile Changes in Coffea spp. Promoted by Single and Combined Exposure to Drought and Elevated CO2 Concentration.

Climate change scenarios pose major threats to many crops worldwide, including coffee. We explored the primary metabolite responses in two Coffea genotypes, C. canephora cv. Conilon Clone 153 and C. arabica cv. Icatu, grown at normal (aCO2) or elevated (eCO2) CO2 concentrations of 380 or 700 ppm, respectively, under well-watered (WW), moderate (MWD), or severe (SWD) water deficit conditions, in order to assess coffee responses to drought and how eCO2 can influence such responses. Primary metabolites were analyzed with a gas chromatography time-of-flight mass spectrometry metabolomics platform (GC-TOF-MS). A total of 48 primary metabolites were identified in both genotypes (23 amino acids and derivatives, 10 organic acids, 11 sugars, and 4 other metabolites), with differences recorded in both genotypes. Increased metabolite levels were observed in CL153 plants under single and combined conditions of aCO2 and drought (MWD and SWD), as opposed to the observed decreased levels under eCO2 in both drought conditions. In contrast, Icatu showed minor differences under MWD, and increased levels (especially amino acids) only under SWD at both CO2 concentration conditions, although with a tendency towards greater increases under eCO2. Altogether, CL153 demonstrated large impact under MWD, and seemed not to benefit from eCO2 in either MWD and SWD, in contrast with Icatu.

Response of Key Metabolites during a UV-A Exposure Time-Series in the Cyanobacterium Chlorogloeopsis fritschii PCC 6912.

Ultraviolet A (UV-A) is the major component of UV radiation reaching the Earth’s surface, causing indirect damage to photosynthetic organisms via the production of reactive oxygen species (ROS). In comparison, UV-B causes both direct damage to biomolecules and indirect damage. UV-B is well studied in cyanobacterial research due to their long evolutionary history and adaptation to high levels of UV, with less work on the effects of UV-A. In this study, the response of key metabolites in Chlorogloeopsis fritschii (C. fritschii) during 48 h of photosynthetically active radiation (PAR, 15 µmol·m−2·s−1) supplemented with UV-A (11 µmol·m−2·s−1) was investigated using gas chromatography- mass spectrometry (GC-MS). Results showed an overall significant increase in metabolite levels up to 24 h of UV-A exposure. Compared with previously reported UV-B (PAR + UV-B) and PAR only results, UV-A showed more similarity compared to PAR only exposure as opposed to supplemented UV-B. The amino acids glutamate, phenylalanine and leucine showed differences in levels between UV (both supplemented UV-A and supplemented UV-B) and PAR only (non-supplemented PAR), hinting to their relevance in UV stress response. The fatty acids, palmitic and stearic acid, showed positive log2 fold-change (FC) in supplemented UV-A and PAR only experiments but negative log2 FC in UV-B, indicating the more harmful effect of UV-B on primary metabolism. Less research has been conducted on UV-A exposure and cyanobacteria, a potential environmental stimuli for the optimisation of metabolites for industrial biotechnology. This study will add to the literature and knowledge on UV-A stress response at the metabolite level in cyanobacteria, especially within the less well-known species C. fritschii.

Diet-Derived Circulating Antioxidants and Risk of Coronary Heart Disease: A Mendelian Randomization Study

BackgroundPreviously, observational studies have identified associations between higher levels of dietary-derived antioxidants and lower risk of coronary heart disease (CHD), whereas randomized clinical trials showed no reduction in CHD risk following antioxidant supplementation. ObjectivesThe purpose of this study was to investigate possible causal associations between dietary-derived circulating antioxidants and primary CHD risk using 2-sample Mendelian randomization (MR). MethodsSingle-nucleotide polymorphisms for circulating antioxidants (vitamins E and C, retinol, β-carotene, and lycopene), assessed as absolute levels and metabolites, were retrieved from the published data and were used as genetic instrumental variables. Summary statistics for gene-CHD associations were obtained from 3 databases: the CARDIoGRAMplusC4D consortium (60,801 cases; 123,504 control subjects), UK Biobank (25,306 cases; 462,011 control subjects), and FinnGen study (7,123 cases; 89,376 control subjects). For each exposure, MR analyses were performed per outcome database and were subsequently meta-analyzed. ResultsAmong an analytic sample of 768,121 individuals (93,230 cases), genetically predicted circulating antioxidants were not causally associated with CHD risk. For absolute antioxidants, the odds ratio for CHD ranged between 0.94 (95% confidence interval [CI]: 0.63 to 1.41) for retinol and 1.03 (95% CI: 0.97 to 1.10) for β-carotene per unit increase in ln-transformed antioxidant values. For metabolites, the odds ratio ranged between 0.93 (95% CI: 0.82 to 1.06) for γ-tocopherol and 1.01 (95% CI: 0.95 to 1.08) for ascorbate per 10-fold increase in metabolite levels. ConclusionsEvidence from our study did not support a protective effect of genetic predisposition to high dietary-derived antioxidant levels on CHD risk. Therefore, it is unlikely that taking antioxidants to increase blood antioxidants levels will have a clinical benefit for the prevention of primary CHD.

Metabolomic insights of the tomato fruits (Solanum lycopersicum L.) cultivated under different supplemental LED lighting and mineral nutrient conditions

Appropriate adjustment of various microclimatic and nutrient conditions can improve crop productivity under greenhouse cultivation. Here, we performed non-targeted profiling to investigate metabolite variation in tomato fruits following different supplementary LED lighting (SL) (red: R; blue: B; and a combination of red and blue light: RB, for 1 h/day and 6 h/day per condition) and mineral nutrient supply (N, K, Mg). The different SL treatments affected metabolite variation in tomato fruits more significantly than mineral nutrients. Importantly, regulating the SL period induced a more distinct metabolite composition in tomato fruits than that induced by varying the SL sources. Tomato fruits cultivated under RB for 6 h/day had a relatively higher content of sugar derivatives, especially sucrose, thus influencing the gustatory characteristics of the fruit. In contrast, fruit from tomatoes cultivated under RB for 1 h/day showed a distinctive increase in the abundance of amino acids, organic acids, and several secondary metabolites, adding to its nutritional quality. Intriguingly, the different mineral supplements elicited discriminant metabolic variation in tomato fruits cultivated under reduced levels of mineral supply; however, the effects were insignificant under enhanced levels of mineral supply. Specifically, reduced level in nitrogen supply resulted in a lower abundance of amino acids, whereas reduced level in potassium supply increased metabolite levels including amino acids, sugars, and fatty acids in tomato fruits. In the present study, we employed a non-targeted metabolomics approach to unravel the effects of microclimatic parameters and mineral nutrients on fruit quality parameters of tomato plants, by which regulating the period of SL and reducing the potassium concentration were suggested to improve different nutritional qualities.

Metabolite and Phytohormone Profiling Illustrates Metabolic Reprogramming as an Escape Strategy of Deepwater Rice during Partially Submerged Stress.

Rice varieties that can survive under submergence conditions respond to flooding either by enhancing internode elongation or by quiescence of shoot elongation. Despite extensive efforts to identify key metabolites triggered by complete submergence of rice possessing SUBMERGENCE 1 (SUB1) locus, metabolic responses of internode elongation of deepwater rice governed by the SNORKEL 1 and 2 genes remain elusive. This study investigated specific metabolomic responses under partial submergence (PS) to deepwater- (C9285) and non-deepwater rice cultivars (Taichung 65 (T65)). In addition, we examined the response in a near-isogenic line (NIL-12) that has a C9285 genomic fragment on chromosome 12 introgressed into the genetic background of T65. Under short-term submergence (0–24 h), metabolite profiles of C9285, NIL-12, and T65 were compared to extract significantly changed metabolites in deepwater rice under PS conditions. Comprehensive metabolite and phytohormone profiling revealed increases in metabolite levels in the glycolysis pathway in NIL-12 plants. Under long-term submergence (0–288 h), we found decreased amino acid levels. These metabolomic changes were opposite when compared to those in flood-tolerant rice with SUB1 locus. Auxin conjugate levels related to stress response decreased in NIL-12 lines relative to T65. Our analysis helped clarify the complex metabolic reprogramming in deepwater rice as an escape strategy.

Myo-inositol mediates the effects of traffic-related air pollution on generalized anxiety symptoms at age 12 years

BackgroundExposure to traffic-related air pollution (TRAP) has been linked to childhood anxiety symptoms. Neuroimaging in patients with anxiety disorders indicate altered neurochemistry. ObjectivesEvaluate the impact of TRAP on brain metabolism and its relation to childhood anxiety symptoms in the Cincinnati Childhood Allergy and Air Pollution Study (CCAAPS). MethodsAdolescents (n = 145) underwent magnetic resonance spectroscopy. Brain metabolites, including myo-inositol, N-acetylaspartate, creatine, choline, glutamate, glutamate plus glutamine, and glutathione were measured in the anterior cingulate cortex. Anxiety symptoms were assessed using the Spence Children's Anxiety Scale. TRAP exposure in early-life, averaged over childhood, and during the 12 months prior to imaging was estimated using a validated land use regression model. Associations between TRAP exposure, brain metabolism, and anxiety symptoms were estimated using linear regression and a bootstrapping approach for testing mediation by brain metabolite levels. ResultsRecent exposure to high levels of TRAP was associated with significant increases in myo-inositol (β = 0.26; 95%CI 0.01, 0.51) compared to low TRAP exposure. Recent elevated TRAP exposure (β = 4.71; 95% CI 0.95, 8.45) and increased myo-inositol levels (β = 2.98; 95% CI 0.43, 5.52) were also significantly associated with increased generalized anxiety symptoms with 12% of the total effect between TRAP and generalized anxiety symptoms being mediated by myo-inositol levels. ConclusionsThis is the first study of children to utilize neuroimaging to link TRAP exposure, metabolite dysregulation in the brain, and generalized anxiety symptoms among otherwise healthy children. TRAP may elicit atypical excitatory neurotransmission and glial inflammatory responses leading to increased metabolite levels and subsequent anxiety symptoms.

NMR-based metabolomics to select a surgical method for treating papillary thyroid carcinoma

OBJECTIVE:This study aims to investigate differences in the metabolomic profiles of patients who received different surgeries for papillary thyroid carcinoma (PTC).METHODS:Two surgical methods, i.e., unilateral and total thyroidectomy, were employed according to different disease conditions. Sera from patients who were treated with levothyroxine sodium tablets before and after surgery was analyzed with a Bruker 500 Hz nuclear magnetic resonance (NMR) spectrometer. Data were analyzed via principal component analysis (PCA) and partial least squares discriminate analysis (PLS-DA) with SIMCA-P+ 11.0 software, and metabolites were obtained and compared. The first and second principal components were selected from PCA, PLS-DA, and orthogonal partial least squares discriminate analysis (OPLS-DA). A p-value less than 0.05 was considered statistically significant.RESULTS:There were significant differences in serum metabolomics before and after surgery. Compared with unilateral thyroidectomy, total thyroidectomy reversed some highly increased metabolite levels (e.g., taurine and betaine). More significant variations in abnormal metabolites were noted after total thyroidectomy than after unilateral thyroidectomy (e.g., alanine, choline, hippurate, and formic acid).CONCLUSIONS:The choice of surgical method for PTC patients should be based not only on the tumor condition but also on the potential consequences of metabolic variations. Total thyroidectomy reversed some increased metabolite levels but led to accumulation of some other metabolites due to the loss of thyroid function; thus, metabolic disturbances caused by thyroid hormone deficiency should be prevented in advance.

Identification of a Classical Mutant in the Industrial Host Aspergillus niger by Systems Genetics: LaeA Is Required for Citric Acid Production and Regulates the Formation of Some Secondary Metabolites.

The asexual filamentous fungus Aspergillus niger is an important industrial cell factory for citric acid production. In this study, we genetically characterized a UV-generated A. niger mutant that was originally isolated as a nonacidifying mutant, which is a desirable trait for industrial enzyme production. Physiological analysis showed that this mutant did not secrete large amounts of citric acid and oxalic acid, thus explaining the nonacidifying phenotype. As traditional complementation approaches to characterize the mutant genotype were unsuccessful, we used bulk segregant analysis in combination with high-throughput genome sequencing to identify the mutation responsible for the nonacidifying phenotype. Since A. niger has no sexual cycle, parasexual genetics was used to generate haploid segregants derived from diploids by loss of whole chromosomes. We found that the nonacidifying phenotype was caused by a point mutation in the laeA gene. LaeA encodes a putative methyltransferase-domain protein, which we show here to be required for citric acid production in an A. niger lab strain (N402) and in other citric acid production strains. The unexpected link between LaeA and citric acid production could provide new insights into the transcriptional control mechanisms related to citric acid production in A. niger. Interestingly, the secondary metabolite profile of a ΔlaeA strain differed from the wild-type strain, showing both decreased and increased metabolite levels, indicating that LaeA is also involved in regulating the production of secondary metabolites. Finally, we show that our systems genetics approach is a powerful tool to identify trait mutations.

Abstract 280: Increased Metabolite Levels of Glycolysis and Pentose Phosphate Pathway in Rabbit Atherosclerotic Arteries and Hypoxic Macrophage

Aims: Inflammation and possibly hypoxia largely affect glucose utilization in atherosclerotic arteries, which could alter many metabolic systems. However, metabolic changes in atherosclerotic plaques remain unknown. The present study aims to identify changes in metabolic systems relative to glucose uptake and hypoxia in rabbit atherosclerotic arteries and cultured macrophages. Methods: Macrophage-rich or smooth muscle cell (SMC)-rich neointima was created by balloon injury in the iliac-femoral arteries of rabbits fed with a 0.5% cholesterol diet or a conventional diet. THP-1 macrophages stimulated with lipopolysaccharides (LPS) and interferon-γ (INFγ) were cultured under normoxic and hypoxic conditions. We evaluated comprehensive arterial and macrophage metabolism by performing metabolomic analyses using capillary electrophoresis-time of flight mass spectrometry. We evaluated glucose uptake and its relationship to vascular hypoxia using 18F-fluorodeoxyglucose (18F-FDG) and pimonidazole, a marker of hypoxia. Results: The levels of many metabolites increased in the iliac-femoral arteries with macrophage-rich neointima, compared with those that were not injured and those with SMC-rich neointima (glycolysis, 4 of 9; pentose phosphate pathway, 4 of 6; tricarboxylic acid cycle, 4 of 6; nucleotides, 10 of 20). The uptake of 18F-FDG in arterial walls measured by autoradiography positively correlated with macrophage- and pimonidazole-immunopositive areas (r = 0.76, and r = 0.59 respectively; n = 69 for both; p < 0.0001). Pimonidazole immunoreactivity was closely localized with the nuclear translocation of hypoxia inducible factor-1α and hexokinase II expression in macrophage-rich neointima. The levels of glycolytic (8 of 8) and pentose phosphate pathway (4 of 6) metabolites increased in LPS and INFγ stimulated macrophages under hypoxic but not normoxic condition. Plasminogen activator inhibitor-1 protein levels in the supernatant were closely associated with metabolic pathways in the macrophages. Conclusion: Infiltrative macrophages in atherosclerotic arteries might affect metabolic systems, and hypoxia but not classical activation might augment glycolytic and pentose phosphate pathways in macrophages.

Increased Metabolite Levels of Glycolysis and Pentose Phosphate Pathway in Rabbit Atherosclerotic Arteries and Hypoxic Macrophage

AimsInflammation and possibly hypoxia largely affect glucose utilization in atherosclerotic arteries, which could alter many metabolic systems. However, metabolic changes in atherosclerotic plaques remain unknown. The present study aims to identify changes in metabolic systems relative to glucose uptake and hypoxia in rabbit atherosclerotic arteries and cultured macrophages.MethodsMacrophage-rich or smooth muscle cell (SMC)-rich neointima was created by balloon injury in the iliac-femoral arteries of rabbits fed with a 0.5% cholesterol diet or a conventional diet. THP-1 macrophages stimulated with lipopolysaccharides (LPS) and interferon-γ (INFγ) were cultured under normoxic and hypoxic conditions. We evaluated comprehensive arterial and macrophage metabolism by performing metabolomic analyses using capillary electrophoresis-time of flight mass spectrometry. We evaluated glucose uptake and its relationship to vascular hypoxia using 18F-fluorodeoxyglucose (18F-FDG) and pimonidazole, a marker of hypoxia.ResultsThe levels of many metabolites increased in the iliac-femoral arteries with macrophage-rich neointima, compared with those that were not injured and those with SMC-rich neointima (glycolysis, 4 of 9; pentose phosphate pathway, 4 of 6; tricarboxylic acid cycle, 4 of 6; nucleotides, 10 of 20). The uptake of 18F-FDG in arterial walls measured by autoradiography positively correlated with macrophage- and pimonidazole-immunopositive areas (r = 0.76, and r = 0.59 respectively; n = 69 for both; p<0.0001). Pimonidazole immunoreactivity was closely localized with the nuclear translocation of hypoxia inducible factor-1α and hexokinase II expression in macrophage-rich neointima. The levels of glycolytic (8 of 8) and pentose phosphate pathway (4 of 6) metabolites increased in LPS and INFγ stimulated macrophages under hypoxic but not normoxic condition. Plasminogen activator inhibitor-1 protein levels in the supernatant were closely associated with metabolic pathways in the macrophages.ConclusionInfiltrative macrophages in atherosclerotic arteries might affect metabolic systems, and hypoxia but not classical activation might augment glycolytic and pentose phosphate pathways in macrophages.

Heat and Cold Stresses Phenotypes of Arabidopsis thaliana Calmodulin Mutants: Regulation of Gamma-Aminobutyric Acid Shunt Pathway under Temperature Stress

Plants have evolved mechanisms to cope with changes in surrounding temperatures. T-DNA insertions in seven calmodulin genes of Arabidopsis thaliana were used to investigate the role of specific calmodulin isoforms in tolerance of plants to low and high temperature for seed germination, susceptibility to low and high temperature induced oxidative damage, and changes in the levels of gammaaminobutyric acid (GABA) shunt metabolites in response to temperature stress. Exposure of wild type (WT) and cam mutant seeds at 4 °C showed reduction in germination of cam5-4 and cam6-1 seeds. Exposure of cam seedlings to 42 °C for 2 h showed reduction in seed germination and survival of seedlings in cam5-4 and cam6-1 mutants compared to WT and other cam mutants. Oxidative damage by heat and cold stress measured as the level of malonaldehyde (MDA) was detected increased in root and shoot tissues of cam5-4 and cam6-1. Oxidative damage by heat measured as the level of MDA was detected in root and shoot of most cam mutants with highest levels in cam5-4 and cam6-1. Level of GABA shunt metabolites in seedlings were gradually increased after 1 h and 3 h with maximum level after 6 h and 12 h treatments at 4 °C. GABA shunt metabolites in both root and shoot were generally elevated after 30 min and 1 h treatment at 42 °C, and increased substantially after 2 h at 42 °C comparing to the control (no treatment). GABA and glutamate levels were increased significantly more than alanine in root and shoot tissues of all cam mutants and wild type compared to the control. Alanine levels showed significant decreases in all cam mutants and in WT for 30 and 60 min of heat stress. Sensitivity of cam5-4 and cam6-1 to low temperatures suggests a role of the CAM5 and CAM6 genes in seed germination and protection against cold induced oxidative damage. Increases in the level of GABA shunt metabolites in response to cold treatment after initial reduction in some cam mutants suggests a role for calmodulin protein (cam) in the activation of glutamate decarboxylase (GAD) after exposure to cold, while increased metabolite levels may indicate involvement of other factors like reduction in cytoplasmic pH in cold regulation. Initial general elevation in GABA shunt metabolites after 30 min heat treatment in cam mutants suggests regulation of GABA level by cam. These data suggest that regulation by factors other than cam is likely, and that this factor may relate to the regulation of GAD by intracellular pH and/or metabolite partitioning under heat stress.

Antipsychotic drug effects on left prefrontal phospholipid metabolism: A follow-up 31P-2D-CSI study of haloperidol and risperidone in acutely ill chronic schizophrenia patients

Introduction31Phosphorous magnetic resonance spectroscopy (2D chemical shift imaging, CSI) allows multiregional study of membrane phospholipids and high-energy phosphates in vivo. Increased membrane lipid turnover and impaired energy supply have repeatedly been shown in first-episode schizophrenia patients, and might be a target of drug actions other than dopamine receptors. Here, we explored differential metabolic effects of a typical vs. an atypical antipsychotic on brain phospholipids. MethodsWe applied 2D-CSI MR spectroscopy in 17 recurrent-episode schizophrenia patients off antipsychotics at baseline and at follow-up after 6weeks, during which 7 patients were treated with haloperidol (10–16mg/d) and 10 with risperidone (4–6mg/d). Psychopathology changes were assessed using PANSS, BPRS and CGI scores. ResultsFollow-up analysis using repeated measure ANOVA revealed different effects of both antipsychotic agents: while risperidone generally increased metabolite levels, haloperidol showed a tendency to decrease them. This diverging effect was significant for ATP levels in the left lateral frontal cortex. Furthermore, risperidone increased ATP in the left dorsolateral prefrontal cortex, left anterior temporal cortex and left insular cortex, basal ganglia, and anterior cerebellum, along with left frontal and prefrontal increase of PCr, PDE and PME in these brain regions. ConclusionRisperidone seems to stimulate neuronal and synaptic phospholipid remodeling in left frontal and prefrontal regions, and to a lesser extent also in temporal and insular cortices. We discuss these effects with respect to clinical effects on negative and cognitive symptoms, as well as interaction of phospholipid metabolism with glutamatergic neurotransmission.